Saha Sathi et al / Int. J. Res. Ayurveda Pharm. 4(2), Mar – Apr 2013
Research Article
www.ijrap.net
PHARMACOGNOSTIC AND ANATOMICAL STUDIES OF ANTIGONON LEPTOPUS HOOK. AND ARN.:
A PROMISING MEDICINAL CLIMBER
Saha Sathi1 and Rahaman Chowdhury Habibur 2*
1
Research Scholar, Department of Botany, Visva-Bharati, Santiniketan, India
2
Associate Professor, Department of Botany, Visva-Bharati, Santiniketan, India
Received on: 07/12/12 Revised on: 22/01/13 Accepted on: 10/02/13
*Corresponding author
E-mail: [email protected] and [email protected]
DOI: 10.7897/2277-4343.04219
Published by Moksha Publishing House. Website www.mokshaph.com
All rights reserved.
ABSTRACT
The evaluation of quality and purity of crude drugs by means of various parameters is the most important aspect of pharmacognosy. The present study
deals with pharmacognostic characterization of Antigonon leptopus Hook. and Arn. of the family Polygonaceae. The tubers of it are used by the tribal
people as tonic and nutritious food. The parameters like micromorphological, anatomical, phytochemical and physical constants have been employed
for the pharmacognostical evaluation of different parts of this plant. The stomata are of anisocytic type and amphistomatic. Stomatal indices are 11.08
and 12.13 in upper and lower surfaces respectively. Palisade ratio is 5.5. Trichomes are of both nonglandular and glandular types. Methanolic extracts
of leaf indicate presence of alkaloids, flavonoids, tannins, saponins, anthraquinone, etc. Ash value and moisture content of the leaves were found to be
20.68% and 76% respectively. This study will provide some diagnostic features by which the crude drug of this plant can easily be identified.
Key Words: Pharmacognostic and anatomical studies, Antigonon leptopus Hook. and Arn.
INTRODUCTION
Plants have been playing a vital role in curing the diseases
and ailments of human being from the time immemorial.
Herbal medicines are gradually being popular for primary
health care of the human’s world-wide because of their
efficacy, easy availability, no or very negligible side
effect and low price. Approximately 2,400–3,000
medicinal plants species are in use in different Indian
systems of medicine including Ayurveda and many of
those plants are constantly being screened for their
biological activity1. Many of the important medicinal
plants in India have pharmacognostically characterized
and they have been enumerated in standard literature on
pharmacognosy2. It is also found that pharmacognostic
studies in lesser known medicinal as well as
ethnomedicinal plants are very meager. In this context,
the present study has been undertaken to evaluate this
lesser known ethnomedicinal plant pharmacognostically
including its morpho-anatomy. The leaves of the
investigated plant have been considered here in this
investigation because leaves are commonly used by the
tribal and common people for curing the diseases. Use of
micromorphology and anatomy is now a recognized tool
in the field of plant systematics3-5. Importance of
epidermal characters in general and those of trichomes in
particular and comparative wood anatomy are widely
employed in taxonomic consideration of angiosperms6-10.
Ontogeny and structure of stomata are now also
considered as an important taxonomic character for many
of the angiospermic taxa11-14. Different members of the
family Polygonaceae have been studied anatomically by
the previous workers with special emphasis on stem and
leaf epidermal micromorphology15-17. Chemical analysis
and biological assays are very important aspects in
pharmacognostic evaluation of medicinal plants18-21. The
detailed pharmacognostic study of this member of the
family Polygonaceae has not yet been done. Therefore, in
this investigation the foliar micromorphology, stem xylem
elements, phytochemical screening and physical
evaluation of this ethnomedicinally important taxon have
been carried out. This investigation will provide some
useful markers for identification of the crude drug
obtained from the investigated climber.
MATERIAL AND METHODS
Plant Material
Antigonon leptopus Hook. and Arn.
Family-Polygonaceae
Herbarium voucher number: S.Saha 49 (Visva- Bharati
Herbarium).
Common English name: Coral Vine.
Botanical characters: A large extensive handsome
climber with woody base and tuberous root. Stem angled,
slightly thickened at the node, finely pubescent. Leaves
alternate, usually 4.5-12 × 2.6-5.5 cm, cordate-ovate or
hastate-ovate, entire or undulate, obtuse or acute,
mucronate, hoary, base cordate; with prominent reticulate
vein. Flowers are small, showy, numerous in axillary and
terminal racemes which terminate into tendrils. Perianth
segments oblong-lanceolate, three outer ones larger,
margin finely ciliate, 0.8-1 cm, and bright pink. Stamens
are 8, filaments basally connate into a short ring. Style 3,
stigma capitate. Fruits are achene, 8 mm long, conical,
rounded at base. Seeds with deep longitudinal grooves.
Flowering and fruiting time: August to January.
Distribution: It is native of South America and is
growing in tropical climate including India.
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Table 1: Stomatal features
Plant taxa
Antigonon
leptopus
Leaf
surface
Upper
Lower
Stomatal type
Strictly anisocytic
Mainly anisocytic, few anomocytic and diacytic
Stomatal
length (mm)
21.77
22.22
Stomatal
width (mm)
16.66
17.29
Stomatal
index (SI)
11.08
12.13
Stomatal
frequency/mm2
212.916
238.437
Table 2: Xylem elements characters
Structure
Vessel Elements
Type
Type of perforation plate
Arrangement of perforation plate
Pit
Arrangement of pit
Tail
Length (mm)
Stem
Simple
Transverse or oblique
Simple
Oblique and alternate
Present
145.75
Breadth (mm)
75.05
Frequency /mm2
Wall thickening
Diameter (µm)
Frequency /mm2
27.53
Root
Simple
Transverse or oblique
Simple
Oblique and alternate
Present
177.336(type-I)
134.42(type-II)
39.51(type-I)
136.64(type-II)
16.66
Spiral
32.95
22.03
Pitted
17.143
25
Ends
Pittation
Septation
Length (mm)
Diameter (mm)
Frequency /mm2
Pointed
Absent
Absent
882.6
19.123
60.80
Pointed
Present
Present
427.55
17.108
62.5
Tracheids
Fibres
Table 3: Microchemical and histochemical tests
Tests/Reagents
Tests for
Nature of changes
Dragendroff’s reagent
Wagner’s reagent
Hager’s solution
Ninhydrin
10% NaOH
Salkowski test
Alkaloids
Alkaloids
Alkaloids
Amino acid
Flavonoids
Steroids and
triterpenoids
Reducing sugars
Glycoside
Orange brown ppt
Orange brown ppt
Yellow ppt
Purple colour
Magenta colour
Reddish-blue and
green florescence
Brick red ppt
Blue colour
Cellulose
Tannins
Benedict’s reagent
Kedde reagent
Iodine solution
10% aqueous potassium
dichromate solution
10% aqueous lead
acetate solution
5% aqueous ferric
chloride solution
1% lead acetate
Borntrager’s test
Degree of changes
(Root)
(Shoot)
+
++
+
++
++
++
+
+
+
+++
+++
++
+++
+++
Blue colour
Yellowish-brown ppt
++
+++
++
+++
Tannins
Yellow ppt
+++
+++
Tannins
Greenish-black colour
+++
+++
++
+
+ = Present
+++
++
Saponins
Anthraquinone
-
White ppt
Pink colour
= Absent ;
Habit and habitat: Perennial terrestrial climber; growing
as common weed along the railway track, on the hedges,
tree top and in fallow lands. Occasionally found in the
gardens as an ornamental plant.
Parts used: Root tuber.
Medicinal uses: Tubers are used as tonic and nutritious
food.
Chemical constituents: Information not available.
For the study of foliar epidermis, leaf samples were
cleared following the Bokhari’s method22. The cleared
leaf samples were then mounted on the slide with a drop
of 10% glycerine and 1% aqueous safranin solution and
observed under compound light microscope. For wood
elements study, the stem pieces were macerated following
Histochemical localization test
(stem part)
Vascular bundle, sclerenchyma.
Epidermis, cortical, xylem, pith.
Cortex, vascular bundle.
Sclerenchyma
Xylem, cortex.
Vascular bundle, pith.
Outer layer of cortex and vascular
bundle.
Epidermis, cortex, vascular bundle.
Cortex, vascular bundle,
Hypodermis.
Outer cortex, sclerenchyma ,
vascular bundle.
Epidermis, cortex, vascular bundle,
pith.
Outer cortical layer.
Vascular bundle.
the standard method23; washed several times, teased with
needles, stained in safranin, mounted on the slides with
10% glycerine and observed under microscope. The
drawings of the foliar micromorphological characters and
stem xylem elements were made with the help of camera
lucida and measurements were taken with standardized
ocular micrometer in each case. Finally, the leaf powder
was extracted (Soxhlet extraction) with 90% methanol
and these extracts were used for different chemical colour
reaction tests for identification of different phytochemical
groups. Physical constants and the UV fluorescence
nature of the powder were studied following the standard
methods19.
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Table 4: UV fluorescence nature of the drug powder
Materials and treatments
In fluorescence light
Shoot powder
Reddish brown
Powder as such (Scraped on filter paper with powder)
Treated with 50% nitric acid
Dark maroon
Treated with 5% potassium hydroxide in water
Brown
Treated with 1N hydrochloric acid
Nevi blue
Treated with dilute 80% sulphuric acid
Reddish brown
Brownish red
Treated with antimony trichloride
Treated with acetone
Reddish orange
Treated with methanol
Lemon yellow
Treated with ethanol
Brownish orange
Root powder
Powder as such (Scraped on filter paper with powder)
Grayish green
Treated with 50% nitric acid
Dark maroon
Treated with 5% potassium hydroxide in water
Blackish green
Blackish green
Treated with 1N hydrochloric acid
Treated with dilute 80% sulphuric acid
Blackish green
Treated with antimony trichloride
Faint greenish
Treated with acetone
Yellow lemon
Treated with methanol
Yellow lemon
Yellow lemon
Treated with ethanol
In ordinary light
Soil colour
Orange
Deep reddish brown
Olive green
Yellowish brown
Faded yellow
Green
Light yellowish green
Green
No change
Orange
Reddish brown
Olive green
Yellowish brown
Cream colour
Cream colour
Cream colour
Straw colour
Figure [a] T.S. of stem, [b] T.S. of petiole, [c] Portion of T.S. of root, [d] Portion of T.S. of tuber, [e] Glandular trichome, [f] Lower epidermal
surface, [g] Vessel element of stem, [h] and [i] Vessel element of root, [j] Portion of fibres of root, [k] A portion of tracheid of root, [l] A
portion of tracheid of stem, [m] Ash of shoot portion, [n] Spot test under UV light (365nm) and [o] Colour reaction test.
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RESULTS
Micromorphology
General description along with measurement of the
epidermal cells, stomata, trichomes, and crystals are given
below.
are very long with pits on the sidewall. Diameter of the
tracheid is 17.143 µm and frequency is 25 /mm2. Fibres
are septate with pointed ends. Pits are present. Size of the
fibre is 427.55 µm x 17.108 µm and its frequency is 62.5
/mm2 (Table 2).
Epidermis: Epidermal cells are irregular in shape and cell
wall outlines are strictly wavy in both the surfaces. The
size of the upper epidermal cell is 43.81 µm x 37.62 µm
and it is 45.25 x 25.11 µm on the lower surface. Cell
frequencies in both upper and lower surfaces are 947.14
/mm2 and1596.92/mm2 respectively. Palisade ratio is 4.36.
Stem Anatomy
The transverse section of the stem is circular in outline
with 5 ridges and 5 grooves. Following tissue
organization from periphery towards the center of the
stem is observed (Figure a).
Stomatal complex: Stomata are present on both surfaces
of the leaf i.e. leaves are amphistomatic. In the upper
surface, stomata are anisocytic type; whereas on the lower
surface they are mainly anisocytic type along with few
diacytic type stomata. Stomatal size in the upper surface
is 21.7 µm x 16.66 µm and in the lower surface, it is
22.218µm x 17.29µm. In the upper stomatal frequency is
212.916 /mm2 and it is 238.437/mm2 in lower surface.
Stomatal index is 11.08 in the upper surface and it is
12.13 in lower surface (Table 1).
Trichomes: Trichomes are of both non-glandular and
glandular types. Non-glandular, uniseriate trichomes are
present on both upper and lower surfaces of the leaf. Size
of this structure is 35.178µm x 10.183µm. Frequency is
41.66 /mm2. Glandular trichomes are multicellular,
stalked and present on the both surfaces. The size of this
structure is 47.509µm x 19.144µm. Their frequency is
20.83/mm2. Trichome index is 2.74.
Crystals: Small, sphaeroidal, crystals of Ca-oxalate are
found in some cells of cortex and pith of the petiole. The
diameter of crystals is 31.25 µm and crystal frequency is
26.04/mm2.
Xylem elements
General description along with measurements of the
xylem elements of stem and root have been presented
below.
Epidermis: Epidermis is uniseriate with compactly
arranged epidermal cells and cuticle is thin here. Both
non-glandular and glandular trichomes are present on
epidermis.
Cortex: The cortex is massive and differentiated into
three distinct zones. The first zone is collenchymatous
hypodermis of two-cell thick, lying just beneath the
epidermal layer. Two layers of chlorenchyma cells are
present beneath the hypodermal layer. Characteristic
single layer continuous starch sheath present just below
the chlorenchyma zone.
Pericycle: It is a wavy, thick layer of sclerenchyma cells
present below the starch sheath. At the grooved region of
the stem, the pericycle layer is four celled in thickness
and it is six celled thick at the ridged zone.
Vascular bundles: They are collateral, conjoint and open
type with phloem and xylem. Vascular bundles are
arranged in two rings. Small vascular bundles are present
at the ridged region and the larger ones are present at the
grooved region of the stem. They are connected by a
secondary vascular cambium ring. In the xylem, few
small vessels are radically arranged.
Pith: At the centre of the stem, massive pith is present.
Cells of it are very large, isodiametric and
parenchymatous with profuse intercellular spaces.
Stem: The vessel elements are with simple, oblique to
transversely placed perforation plate. Pits are simple.
Tails are present in some of the vessel element. Size
is145.75 µm x 75.05 µm and frequency is 60.80/mm2.
Tracheids are with spiral sidewall thickening. Diameter of
the tracheid is 32.95µm and frequency is 22.03 /mm2.
Fibres are typically libriform type with pointed ends.
Septa and pits are completely absent. Size of the fibre is
882.6 µm x 19.123 µm; frequency is 27.53 /mm2 (Table
2).
Petiole anatomy: The outline of petiole in transverse
section is horse-shoe shaped. Epidermis is single layered
and cells are compactly arranged. Cuticle is thin. Nonglandular trichomes are present in the epidermis. 3-4
layers of collenchyma cells present just below the
epidermal layer. Altogether 7 vascular bundles are
arranged in ‘U’ shaped manner. A comparatively larger
bundle is present below the open end of ‘U’-shaped
arrangement. Those 7 bundles are arranged in alternate
manner of one smaller bundle with larger one . At the
centre large parenchyma cells with intercellular spaces
forming pith (Figure b).
Root: The vessel elements here are of two types - one
type is very long (39.51 µm x 177.336 µm) with narrow
width and another type is very short (136.64µm x
134.42µm) with greater width. Frequency of the vessel
elements is 16.66/mm2. Perforation plates of the vessel
elements are simple, oblique to transversely place. Here
pits are simple and arranged both in opposite and alternate
rows. Tails are found in some of the elements. Tracheids
Root anatomy: The transverse section of root is more or
less circular in outline and shows following features from
periphery to its centre. A few cell thick cork layer present
at the periphery. Cork cells are polygonal, thick walled
without any intercellular spaces. A narrow cortical zone
present just below the cork layer consisting of smaller
sized parenchymatous cells. Below the cortical zone, a
multiseriate layer of very thick walled sclerenchymatous
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zone is present that is dotted with 1-2 vessels at the
periphery. Solid mass of primary xylem located at the
centre. Primary xylem consists of many metaxylem and
protoxylem elements which are showing the mixed type
arrangement. Above primary xylem cambium cells are
present in discontinuous ring (Figure c).
Tuber anatomy: The transverse section of a tuber shows
a few cell thick cork layer present at the periphery. Cork
cambium layer present just below the cork layer. Patches
of sclerenchyma present in the cortical zone. Size of
parenchymatous cells is gradually increasing towards the
centre and central parenchymatous cells are very large
which contain many storage substances. Intercellular
spaces are profuse here among the parenchyma cells.
Many small vascular patches are observed in this storage
tissue mass of parenchyma (Figure d).
Microchemical evaluation of the powdered drug
Through the phytochemical tests of the methanolic
extracts of shoot and root parts, some important
phytochemical groups have been detected in both the
cases which confirm the medicinal properties of this
plant. In the shoot extract, the detected phytochemical
groups are tannins, alkaloids, flavonoids, reducing sugars,
saponins, glycosides, anthraquinone, etc. Among these
phytochemical groups tannins are present in higher
amount. The phytochemical groups detected in shoot part
are found in root also. In root, the concentration of
tannins, alkaloids, flavonoids, saponins, glycosides is
higher than shoot (Table 3).
Histochemical Study
Histochemical study of the stem indicates presence of
various phytochemical groups localized in different tissue
zones of it. Vascular bundles and cortical zone of the stem
are the sites where comparatively a large number of
phytochemical groups have been observed than the other
tissue zones of the stem. Chemical groups found in these
tissue zones are alkaloids, tannins, glycosides, saponins,
flavonoids, etc. (Table 3).
Physical Evaluation
Physical constant
Ash Value
· Total ash - 20.68%
· Water soluble ash – 46.42%
· Acid soluble ash – 81.48%
· Moisture Content - 76 % (in fresh form).
Fluorescence analysis
The drug powder treated with different chemical reagents
gives characteristic colour when seen under UV light and
it is compared with the colour observed under ordinary
light. In presence of acetone, methanol and ethanol the
powder drugs in ordinary light showed green, olive green
and green colour respectively; but under UV light (365
nm) the same drug samples
gave prominent bright
reddish orange, lemon yellow and brownish orange
permanent fluorescence colour respectively which are
quite distinct from the colour of ordinary light (Table 4).
DISCUSSION
The present study reveals that foliar epidermal features,
stem and root xylem element characters, primary
phytochemical screenings and physical evaluation are of
some importance in identification of this investigated
plant species in its fresh as well as dried form. Some of
the general anatomical characters of the investigated plant
conformed to the features identified in the other members
of the family Polygonaceae earlier by different
workers15,16.
Stomata of investigated plant are mainly of anisocytic
type and sometime very rarely diacytic stomata are
present on lower surface of the leaf. Studies in stomata
can have a great taxonomic as well as pharmacognostic
value in proper identification of different plant taxa
including medicinal plants11,12,19. In this plant, the
differences in stomatal size and frequency on both the
surfaces have been found. Stomatal frequency of the
lower surface is higher (238.437/ mm2) than the upper
surface. There is a marked difference between stomatal
indices of upper (11.08%) and lower (12.13%) surfaces of
the leaf which is a distinct feature of this plant.
Trichome features are also very important in proper
identification of the plants and considered as one of the
valuable taxonomic markers now24. Epidermal trichomes
of the investigated plant are non-glandular and glandular
types.
Trichome indices of the non-glandular and
glandular trichomes are 20.83% and 41.66% respectively.
Vessel elements of root of the investigated plant show a
characteristic feature in respect of their size. One group of
element is long and narrow and another one is very wide
and short. Septate fibres are observed in the root which
indicates a distinct feature for identification of it. The
arrangement and number of vascular bundles in the
petiole may sometimes provide the diagnostic feature for
identification of the plant species. Here in the petiole,
altogether 7 vascular bundles are found and they are
arranged in ‘U’ shaped manner which can be used as a
marker for identification of this species of Polygonaceae.
Chemical analysis and biological assays are considered as
important aspects in pharmacognostic evaluation of the
medicinal plants19,21. Through phytochemical tests of
methanolic shoot extract of this plant, it is found that the
important phytochemical groups like alkaloids,
glycosides, flavonoids, anthraquinone, tannins, saponins,
etc. are present in this plant which confirms its medicinal
properties. Another phytochemical study on flowers of
Antigonon leptopus revealed that presence of volatile oil,
carboxylic acid, glycoside, carbohydrate, terpenes25. It
corresponds with some of the phytochemical groups
observed in present study. The physical constants like ash
value (20.68%), moisture content (76%) and UV
fluorescence characters of the powder drug of this plant
can also be used as important characters for proper
identification of crude drug of it. Finally some diagnostic
features have been given below which can be employed
for easy identification of Antigonon leptopus in its fresh
as well as dried form.
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Diagnostic features
1. Stomatal indices of upper and lower surfaces are 11.08
and 12.13 respectively.
2. In petiole, number of vascular bundles is 7 and
arranged in ‘U’ shaped manner.
3. Septate fibres are present in the root.
4. Ash Value- total ash - 20.68%
ACKNOWLEDGEMENT
We are very grateful to the Head, Department of Botany, Visva-Bharati,
for providing the necessary laboratory facilities.
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Cite this article as:
Saha Sathi and Rahaman Chowdhury Habibur. Pharmacognostic and
anatomical studies of Antigonon leptopus Hook. and Arn.: A promising
medicinal climber. Int. J. Res. Ayurveda Pharm. 2013; 4(2):186-191
Source of support: Nil, Conflict of interest: None Declared
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